鲤鱼肌肉ACE抑制肽制备及稳定性的研究
发布时间：2018-10-16 11:59
【摘要】：血管紧张素转换酶抑制肽(Angiotensin-I-converting enzyme inhibitory peptide,ACEIP)可以通过与ACE结合而起到调节血压的作用,鲤鱼(Cyprinus carpio;common carp)是一种蛋白质含量较高的动物,我国目前是全世界鲤鱼产量最高的国家,由于加工技术落后,我国的鲤鱼多数用于生鲜或者冷冻,导致鲤鱼的附加值降低,蛋白质资源浪费。本课题研究了从鲤鱼肌肉蛋白中得到ACEIP的提取工艺、分离纯化步骤、氨基酸序列的鉴定以及其稳定性,主要结论如下:1.本课题以鲤鱼肌肉为原料,ACE抑制率为指标,不同蛋白酶酶处理鲤鱼肌肉上清液的ACE抑制活性大小排序如下:碱性蛋白酶中性蛋白酶胰蛋白酶木瓜蛋白酶≈胃蛋白酶;碱性蛋白酶处理鲤鱼肌肉上清液ACE抑制率为86.0±2.29%,明显高于其他蛋白酶,因此选取碱性蛋白酶为提取提取鲤鱼肌肉ACEIP的工具酶。2.采用三因素三水平响应面实验设计,研究了加酶量、酶解温度、酶解时间对于碱性蛋白酶酶解鲤鱼肌肉制备ACEIP的提取工艺,响应面优化法得到最优工艺条件为:加酶量为:4.32U/mg、酶解时间:4h,酶解温度:50℃,鲤鱼肌肉酶解上清液的ACE抑制率达到93.26±1.70%。3.对ACE抑制率测定反应体系进行调整,在最优提取工艺条件下,对酶解上清液进行了 3KDa超滤膜超滤,得到分子量3KDa组分(36.03±2.55%)和分子量3KDa(55.31±1.66%)的两部分,对比得到分子量较少的组分ACE抑制活性更高。4.利用Rp-HPLC法对酶解液进一步分离纯化,筛选每次分离过程中ACE抑制活性最高的组分,最终得到纯度为95.77%的组分P-4,通过N-端测序确定P-4的氨基酸序列为 Tyr-Leu-Lys(YLK)。5.YLK对于ACE的稳定性结果表明,YLK属于抑制剂类型肽;热稳定性、酸碱稳定性、胃肠道消化稳定性研究结果表明YLK对于热处理,酸碱处理,胃肠道消化酶都不敏感,具有很好的加工适应性。
[Abstract]:Angiotensin converting enzyme inhibitory peptide (Angiotensin-I-converting enzyme inhibitory peptide,ACEIP) can regulate blood pressure by binding with ACE. Carp (Cyprinus carpio;common carp) is a kind of animal with high protein content. Due to the backward processing technology, most carp in our country are used for fresh or frozen, which leads to the reduction of the added value of carp and the waste of protein resources. In this paper, the extraction process of ACEIP from carp muscle protein, the steps of separation and purification, the identification of amino acid sequence and its stability were studied. The main conclusions are as follows: 1. Using carp muscle as raw material and ACE inhibition rate as index, the order of ACE inhibitory activity of the supernatant of carp muscle treated with different protease enzymes is as follows: alkaline protease neutral protease trypsin papain 鈮，
本文编号：2274287
[Abstract]:Angiotensin converting enzyme inhibitory peptide (Angiotensin-I-converting enzyme inhibitory peptide,ACEIP) can regulate blood pressure by binding with ACE. Carp (Cyprinus carpio;common carp) is a kind of animal with high protein content. Due to the backward processing technology, most carp in our country are used for fresh or frozen, which leads to the reduction of the added value of carp and the waste of protein resources. In this paper, the extraction process of ACEIP from carp muscle protein, the steps of separation and purification, the identification of amino acid sequence and its stability were studied. The main conclusions are as follows: 1. Using carp muscle as raw material and ACE inhibition rate as index, the order of ACE inhibitory activity of the supernatant of carp muscle treated with different protease enzymes is as follows: alkaline protease neutral protease trypsin papain 鈮，
本文编号：2274287
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